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Nucleic Acids Research, 1986, Vol. 14, No. 14 5693-5711
© 1986

Articles

Mechanism of autonomous control of the Escherichia coli F plasmid: different complexes of the initiator/repressor protein are bound to its operator and to an F plasmid replication origin

Luke Masson and Dan S. Ray*

Department of Biology and Molecular Biology Institute, University of California Los Angeles, CA 90024, USA

*To whom correspondence should be addressed

Received May 8, 1986. Revised June 26, 1986. Accepted June 26, 1986.

E protein, the 29 kd product of the F plasmid repE gene, plays both positive and negative roles in the autoregulation of F replication. We have cloned and expressed the repE gene in an inducible ATG-fusion vector and have detected specific binding of E protein to the repE operator and to four 19-base pair direct repeats (incB) within the F plasmid replication origin ori2 Binding of E protein at the repE operator occurs with higher affinity than at ori2(incB) and gives almost complete protection to at least 30 base pairs, whereas binding of E protein to the direct repeats in the ori2 region shows an alternating pattern of enhanced and reduced sensitivity to DNAase cleavage consistent with a protein-induced folding of the DNA. These results provide direct biochanical support for a model of F plasmid replication in which the E protein serves both as an initiator of replication and as an autorepressor of its own synthesis.


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